Hymenochaetaceae or inonotus obliquus compositions and therapeutic uses thereof

Abstract

The present invention relates to a composition comprising an extract of a fungus belonging to the family Hymenochaetaceae, for use in the treatment or prevention of a pathological condition of the central and / or peripheral nervous system. Preferably, the fungus belonging to the family Hymenochaetaceae is Inonotus obliquus (Chaga). Furthermore, the invention relates to a food supplement comprising the Chaga mushroom.

Description

[0001] HYMENOCHAETACEAE OR INONOTUS OBLIQUUS COMPOSITIONS AND THERAPEUTIC USES THEREOF

[0002] Field of the art

[0003] The present invention relates to a composition comprising an extract of a fungus for the treatment or prevention of a pathological condition of the central and / or peripheral nervous system.

[0004] Prior art

[0005] A-lipoic acid (ALA) is a component produced by plants, animals and human beings. ALA possesses a great antioxidant potential and is widely used as a racemic drug for pain and paraesthesia associated with diabetic polyneuropathy. However, the therapeutic efficacy of ALA is relatively low due to its pharmacokinetic profile; in fact, the data suggest that ALA has a short half-life and limited bioavailability (about 30%) as a result of its degradation in the liver, low solubility and instability in the stomach. It has been observed that ALA is capable of protecting hippocampal neurons against neurotoxicity induced by beta-amyloid. Furthermore, ALA has shown to be important also in countering inflammation and pain. Numerous clinical studies which investigated the properties of ALA in the treatment of pain in neuropathies, the treatment of Alzheimer’s and the treatment of dementia and other cognitive disorders arising as a result of HIV infection have been reported in the literature.

[0006] Due to the regulatory status of food supplements, including the fact that they are not included in pharmacovigilance systems, their safety profile has not been thoroughly studied, despite the growing interest in effectiveness, as demonstrated by ongoing clinical studies. The identification of potential risks is particularly difficult due to the complexity of notification of harmful events tied to food supplements: although in the United States the Center for Food Safety and Applied Nutrition (CFSAN) and Adverse Event Reporting System (CAERS) collect complaints and AR data on products for foods, supplements and cosmetics, in Europe there is no uniformity as to how to report toxicities associated with food supplements. In particular, besides the numerous cases of acute intoxication and insulin autoimmune syndrome (also known as Hirata’s disease), the assessment of the safety of food supplements containing ALA seems to have been neglected. In fact, it has been discovered that many ALA-based supplements have a percentage of serious and unpredictable adverse reactions in Italy, but also at a global level. The early onset of skin and immune disorders, together with the rare occurrence of cholestatic hepatitis, requires a careful assessment of the risk-benefit ratio and continuous monitoring by all stakeholders, including prescribers, consumers, regulators and researchers.

[0007] The present treatments for these conditions often show limitations in terms of effectiveness and side effects, leading researchers to explore alternative therapeutic approaches.

[0008] Food supplements and nutraceuticals have gained popularity as potential complementary or alternative options to support the health of the nervous system. However, many existing products lack sufficient scientific evidence to support the stated benefits, and there is thus a need for new, carefully studied ingredients that may effectively address the complex nature of neurological and neurodegenerative disorders.

[0009] Summary of the invention

[0010] A first aspect of the present invention relates to a composition comprising an extract of a fungus belonging to the family Hymenochaetaceae, for use in the treatment or prevention of a pathological condition of the central and / or peripheral nervous system. Preferably, the fungus belonging to the family Hymenochaetaceae is Inonotus obliquus (Chaga).

[0011] Preferably, the composition further comprises at least one vitamin and / or an amino acid derivative, more preferably chosen from: Vitamin B1, Vitamin B2, Vitamin B6, Vitamin E, Vitamin C, Vitamin B9, Vitamin B12 and Vitamin B5 and the at least one amino acid derivative is carnitine. Preferably, the composition further comprises at least one precursor of the synthesis of membrane phospholipids, more preferably citicoline.

[0012] In one embodiment, the composition is used for the prevention or treatment of a pathological condition selected from the group consisting of Alzheimer’s disease, Parkinson’s disease, multiple sclerosis, neuropathic pain, epilepsy and stroke.

[0013] In a second aspect, the invention relates to a food supplement comprising an extract of a fungus belonging to the family Hymenochaetaceae, at least one vitamin, at least one amino acid derivative and at least one precursor of the synthesis of membrane phospholipids.

[0014] Preferably, the fungus belonging to the family Hymenochaetaceae is Inonotus obliquus (Chaga).

[0015] In one embodiment, the at least one vitamin is chosen from: Vitamin B1, Vitamin B2, Vitamin B6, Vitamin E, Vitamin C, Vitamin B9, Vitamin B12, Vitamin B5, the at least one amino acid derivative is carnitine and the at least one precursor of the synthesis of membrane phospholipids is citicoline.

[0016] Brief description of the drawings

[0017] The present invention will be described in detail below and exemplified, by way of non-limiting demonstration, also with the aid of the appended Figures.

[0018] Figure 1 shows graphs of the survival test on intestinal cells carried out using a Chaga extract, carnitine or citicoline or two compositions comprising a Chaga extract, citicoline and carnitine. The data are expressed as the mean ± SD (%) of 5 independent experiments normalised to the control. p<0.05 vs control; * p<0.05 vs single agents; # p<0.05 vs Mix 1.

[0019] Figure 2 shows graphs of the intestinal permeability test carried out using a Chaga extract, carnitine or citicoline or two compositions comprising a Chaga extract, citicoline and carnitine. The data are expressed as the mean ± SD (%) of 5 independent experiments normalised to the control. p<0.05 vs control; * p<0.05 vs single agents.

[0020] Figure 3 shows the analysis of tight junctions, namely Claudin (A); Occludin (B) and Zo-1 (C).

[0021] The data are expressed as the mean ± SD (%) of 5 independent experiments normalised to the control.

[0022] * p<0.05 vs control; bars p<0.05 vs single agents; a p<0.05 vs Mix 1.

[0023] Figure 4 shows the analysis of safety at the level of the central nervous system. Cell survival (A) and ROS production (B) were assessed. The data are expressed as the mean ± SD (%) of 5 independent experiments normalised to the control. * p<0.05 vs control; bars p<0.05 vs single agents; a p<0.05 vs Mix 1 ; p<0.05 vs H2O2.

[0024] Figure 5 shows the analysis of safety at the level of the central nervous system. The expression of the genes SIRT-1 (A), pTau (B) and APP (C) was assessed. The data are expressed as the mean ± SD (%) of 5 independent experiments normalised to the control. * p<0.05 vs control; bars p<0.05 vs single agents; a p<0.05 vs Mix 1 ; p<0.05 vs H2O2.

[0025] Figure 6 shows the analysis of safety at the level of the peripheral nervous system. Cell survival (A) and ROS production (B) were assessed. The data are expressed as the mean ± SD (%) of 5 independent experiments normalised to the control. * p<0.05 vs control; bars p<0.05 vs single agents; a p<0.05 vs Mix 1 ; 0 p<0.05 vs GGF 200 ng / mL.

[0026] Figure 7 shows the analysis of markers, namely p75 (A) and MPZ (B), at the level of the peripheral nervous system. The data are expressed as the mean ± SD (%) of 5 independent experiments normalised to the control. * p<0.05 vs control; bars p<0.05 vs single agents; a p<0.05 vs Mix 1 ; 0 p<0.05 vs GGF 200 ng / mL.

[0027] Figure 8 shows the analysis of markers, namely NRG1 (A) and ERb (B), at the level of the peripheral nervous system. The data are expressed as the mean ± SD (%) of 5 independent experiments normalised to the control. * p<0.05 vs control; bars p<0.05 vs single agents; a p<0.05 vs Mix 1 ; p<0.05 vs GGF 200 ng / mL.

[0028] Figure 9 shows an assessment of systemic effects of the administration of the single compounds and of a mixture comprising them. The % of triglycerides (A) and total cholesterol (B) were determined.

[0029] Detailed description of preferred embodiments of the invention

[0030] A first aspect of the present invention relates to a composition for use in the treatment or prevention of a pathological condition of the central and / or peripheral nervous system.

[0031] In one embodiment, the composition comprises an extract of Inonotus obliquus, commonly known as the Chaga mushroom. This extract can be obtained from the fruiting body of the Chaga mushroom, which mainly grows on birches in cold climates.

[0032] Preferably, the composition comprises the Inonotus obliquus extract in a percentage by weight of between 15 and 85% weight / weight (w / w) relative to the weight of the composition, preferably between 20 and 80% w / w, even more preferably between 30 and 70% w / w.

[0033] The Chaga mushroom extract can be prepared with various methods. In some cases, the Chaga mushroom is dried and ground until a fine powder is obtained. This ground material can then be extracted with a suitable solvent, such as water, ethanol, methanol or mixtures thereof. The extraction process can facilitate the release of bioactive compounds from the fungal material in the solvent, resulting in an extract which contains these compounds.

[0034] In a preferred embodiment of the invention, the Chaga mushroom extract comprises a quantity of beta-glucans of between 15 and 55% weight / weight of the extract, preferably between 20 and 50% w / w, even more preferably between 30 and 45% w / w. Preferably, the Chaga mushroom extract comprises a quantity of glucans greater than 40% by weight. In one embodiment, the Chaga mushroom extract is obtained by solvent extraction. Preferably, the extract is obtained from the fruiting body or from at least a portion of the fruiting body of the mushroom, is dried and placed in contact, preferably by immersion, in at least one solvent in order to obtain the extract.

[0035] Preferably, the solvent is an aqueous solvent, more preferably water, or it is an alcoholic solvent, more preferably ethanol, or else it is a hydroalcoholic mixture of water and ethanol. Preferably, the solvent is aqueous, more preferably it is water.

[0036] In one embodiment, the fruiting body is subjected to extraction in water, preferably at a temperature of between 80 and 100°C, preferably for a time of between 1 and 10 hours, more preferably between 2 and 6 hours. In one embodiment, the extract is subjected to at least one subsequent concentration or drying or lyophilisation step. The ratio between the solvent and solid material may vary depending on the degree of grinding and the desired yield, with typical values ranging between 1 :10 and 1 :30 g / mL (solid / liquid). In one embodiment, at the end of the treatment, the insoluble residue is separated by filtration or centrifugation, and the extract is concentrated and subjected to precipitation with ethanol in order to obtain the polysaccharide fraction.

[0037] In a further embodiment, the Chaga mushroom extract is obtained with an extraction by means of organic solvents, preferably ethanol or methanol. The extraction can take place at room temperature or under reflux conditions, with optional subsequent fractionation steps to separate the components based on their solubility. These procedures make it possible to obtain extracts with a selective content of lipophilic or aromatic compounds.

[0038] In another embodiment, the Chaga mushroom extract is obtained through ultrasonic extraction, preferably using high-frequency sound waves, more preferably higher than 40 kHz, in order to break the cell walls and facilitate extraction. In one embodiment, the extract is subjected to at least one subsequent concentration or drying or lyophilisation step.

[0039] In another embodiment, the Chaga mushroom extract is obtained by means of microwave-assisted extraction. The application of microwave radiation enables rapid heating of the system and extraction of the compounds in times of less than 60 minutes. In one embodiment, the extract is subjected to at least one subsequent concentration or drying or lyophilisation step.

[0040] In another embodiment, the Chaga mushroom extract is obtained by means of enzymatic extraction, preferably using a mixture of enzymes containing cellulase and pectinase. In one embodiment, the extract is subjected to at least one subsequent concentration or drying or lyophilisation step.

[0041] Irrespective of the type of extraction process, at the end of the extraction step, the raw extracts of Inonotus obliquus are subjected to purification processes aimed at removing impurities and separating the various bioactive fractions. In general, the purification begins with the concentration of the aqueous extract by vacuum evaporation or lyophilisation, followed by one or more selective precipitations in organic solvents, typically ethanol, in concentrations of between 70% and 95%. This step allows for the separation of the polysaccharide fraction from the low molecular weight components and protein residues.

[0042] The precipitated fraction can be further treated by dialysis or ultrafiltration to remove salts, simple sugars and residual solvents. Alternatively, chromatography techniques, such as ion exchange or gel filtration chromatography, may be used to obtain polysaccharides or triterpenoids with a controlled degree of purity and defined molecular weight.

[0043] For the phenolic and triterpenoid fractions, the purification can include liquid-liquid extraction with solvents of different polarity or normal-phase or reverse-phase column purification. In some cases, the combination of concentration, fractionation and chromatography makes it possible to obtain compounds with a purity greater than 90%. In conclusion, the purification steps can be adapted based on the composition of the initial extract and the intended use of the final product, while maintaining the chemical stability of the compounds and reducing the loss of biological activity to a minimum.

[0044] In some cases, the Chaga mushroom extract is standardised to ensure a predetermined concentration of at least one active compound. The standardisation may entail the adjustment of the concentration of the active compound in the extract to make it fall within a predetermined range. This can be obtained, for example, using analysis by high-performance liquid chromatography (HPLC) to determine the concentration of the active compound in the extract, and then adjusting the concentration according to need.

[0045] The active compounds in the Chaga mushroom extract can include, for example, betulinic acid, betulin, inotodiol, trametenolic acid and ergosterol peroxide. These compounds are known for their potential therapeutic effects, and their presence in the extract can contribute to the potential effectiveness of the composition in the treatment or prevention of pathological conditions of the central and / or peripheral nervous system. In one embodiment, the composition comprising the Chaga mushroom extract is used in the treatment or prevention of a disease of the central and / or peripheral nervous system, preferably selected from Alzheimer’s disease, Parkinson’s disease, multiple sclerosis, neuropathic pain, epilepsy and stroke. Preferably, the composition is administered as a tablet, capsule, liquid or powder, and can be taken by mouth or administered through other suitable routes.

[0046] In some embodiments, the process of standardisation can entail the concentration of the extract by evaporation or lyophilisation. This can serve to increase the concentration of the active compounds in the extract, thus ensuring that the final composition contains a predetermined concentration of these compounds.

[0047] In one embodiment, the solvent used for the extraction process can be selected from the group consisting of water, ethanol, methanol and mixtures thereof. The choice of the solvent may depend on various factors, such as the desired concentration of the active compounds in the extract, the solubility of the active compounds in the solvent and the intended use of the extract.

[0048] In one embodiment, the composition further comprises at least one vitamin. The inclusion of vitamins in the composition can provide additional benefits, since vitamins are known to play crucial roles in various physiological processes, including those tied to the functioning of the nervous system. The specific vitamin or combination of vitamins included in the composition may depend on the intended use of the composition. In one embodiment, the composition comprises at least one vitamin chosen from a group consisting of: Vitamin B1, Vitamin B2, Vitamin B6, Vitamin E, Vitamin C, Vitamin B9, Vitamin B12 and Vitamin B5.

[0049] Preferably, the composition comprises Vitamin B1, also known as thiamine, which is essential for the metabolism of carbohydrates and energy production in the body. Thiamine also plays a role in nerve function and can contribute to the health of the nervous system.

[0050] Preferably, the composition comprises Vitamin B2, or riboflavin, which is involved in energy production and in the metabolism of fats, ketone bodies, carbohydrates and proteins. Riboflavin is also important for maintaining the body’s antioxidant system.

[0051] Preferably, the composition comprises Vitamin B6, also known as pyridoxine, which is involved in the synthesis of neurotransmitters, the chemical messengers that transmit signals in the nervous system. Pyridoxine also plays a role in the production of red blood cells and in protein metabolism.

[0052] Preferably, the composition can also include Vitamin E, a liposoluble vitamin that acts as an antioxidant, protecting cells against damage caused by free radicals. Vitamin E also plays a role in immune function and in DNA repair. Preferably, the composition comprises Vitamin C, also known as ascorbic acid, which is a powerful antioxidant capable of neutralising harmful free radicals. In addition to its antioxidant function, Vitamin C is also involved in the synthesis of collagen, a protein that imparts structure to the skin, hair, nails, blood vessels and bones.

[0053] Preferably, the composition comprises Vitamin B9, also known as folic acid or folate, which is essential for the production and maintenance of new cells. Folic acid is also involved in DNA and RNA synthesis and in amino acid metabolism.

[0054] Preferably, the composition comprises Vitamin B12, also known as cobalamin, which is involved in the metabolism of every cell of the human body. Cobalamin is particularly important in the normal functioning of the nervous system and in DNA and RNA synthesis.

[0055] Preferably, the composition comprises Vitamin B5, also known as pantothenic acid, which is involved in the synthesis and metabolism of proteins, carbohydrates and fats. Pantothenic acid also plays a role in the production of coenzyme A, a molecule involved in numerous metabolic pathways.

[0056] In one embodiment, the composition comprises a combination of these vitamins. The specific combination of vitamins included in the composition can be selected based on the intended use of the composition and the potential benefits of the vitamins.

[0057] In a preferred embodiment, the composition comprises at least two, or at least three, or at least four, or all the vitamins of the group consisting of: Vitamin B1, Vitamin B2, Vitamin B6, Vitamin E, Vitamin C, Vitamin B9, Vitamin B12 and Vitamin B5.

[0058] In one embodiment, the composition further comprises at least one amino acid derivative. Amino acid derivatives are compounds derived from amino acids and can have various physiological effects. The specific amino acid derivative or the combination of amino acid derivatives included in the composition can depend on the intended use of the composition. Preferably, the amino acid derivative included in the composition is carnitine. Carnitine is a quaternary ammonium compound involved in the transport of fatty acids in mitochondria, where they are degraded through beta-oxidation to generate adenosine triphosphate (ATP), the main carrier of energy in cells. Carnitine can also have antioxidant properties and can contribute to the health of the nervous system.

[0059] In some aspects, the inclusion of carnitine in the composition can provide additional benefits. For example, carnitine can improve the production of energy in neurons, support nervous system function and contribute to the potential therapeutic effects of the composition in the treatment or prevention of pathological conditions of the central and / or peripheral nervous system.

[0060] In one embodiment, the composition further comprises at least one precursor of the synthesis of membrane phospholipids. Precursors of the synthesis of membrane phospholipids are compounds involved in the synthesis of phospholipids, the main components of cell membranes. The inclusion of such precursors in the composition can contribute to nervous system health and function, since phospholipids play crucial roles in the structure and function of neurons, the cells making up the nervous system. Preferably, the precursor of the synthesis of membrane phospholipids included in the composition is citicoline. Citicoline is a compound involved in the synthesis of phosphatidylcholine, an important phospholipid in cell membranes. Citicoline can contribute to the health of neurons by supporting the synthesis of phosphatidylcholine, thus promoting the integrity and function of neuronal cell membranes.

[0061] The inclusion of citicoline in the composition can provide additional benefits. For example, citicoline can support nervous system function, improve neuron repair and contribute to the potential therapeutic effects of the composition in the treatment or prevention of pathological conditions of the central and / or peripheral nervous system.

[0062] In a preferred embodiment, the composition comprises or consists of Vitamin B1, Vitamin B2, Vitamin B6, Vitamin E, Vitamin C, Vitamin B9, Vitamin B12 and Vitamin B5, carnitine, citicoline and the Chaga mushroom extract.

[0063] Preferably, the composition can be used in the treatment or prevention of a disease of the central and / or peripheral nervous system. This disease is preferably selected from: Alzheimer's disease, Parkinson's disease, multiple sclerosis, neuropathic pain, epilepsy and stroke.

[0064] In some aspects, specific formulations of the composition can be developed for targeted applications in the treatment or prevention of nervous system disorders. For example, one formulation can comprise an extract of the fungus Inonotus obliquus (Chaga) standardised to a predetermined concentration of betulinic acid, together with Vitamin B1 and carnitine. This formulation can be particularly beneficial for conditions such as Alzheimer’s disease, where the potential therapeutic effects of betulinic acid, the role of Vitamin B1 in the metabolism of carbohydrates and in energy production, and the involvement of carnitine in energy production in neurons can collectively contribute to nervous system health and function.

[0065] In another example, one formulation can comprise an Inonotus obliquus extract standardised to a predetermined concentration of inotodiol, together with Vitamin B6 and citicoline. This formulation can be beneficial for conditions such as Parkinson’s disease, where the potential therapeutic effects of inotodiol, the role of Vitamin B6 in the synthesis of neurotransmitters, and involvement of citicoline in the synthesis of phosphatidylcholine can collectively support nervous system health and function.

[0066] In yet another example, one formulation can comprise an Inonotus obliquus extract standardised to a predetermined concentration of ergosterol peroxide, together with Vitamin E and carnitine. This formulation can be beneficial for conditions such as multiple sclerosis, where the potential therapeutic effects of ergosterol peroxide, the antioxidant properties of Vitamin E, and the role of carnitine in energy production in neurons can collectively contribute to nervous system health and function. A second aspect of the present invention relates to a food supplement comprising the composition described above in detail. This food supplement can comprise an extract of the fungus Inonotus obliquus (Chaga), which can be standardised to ensure a predetermined concentration of at least one active compound. The active compound can be selected from a group consisting of betulinic acid, betulin, inotodiol, trametenolic acid and ergosterol peroxide.

[0067] Preferably, in addition to the Chaga extract, the food supplement further comprises at least one vitamin, at least one amino acid derivative and at least one precursor of the synthesis of membrane phospholipids. The inclusion of these additional components can provide a complete approach to support nervous system health and function.

[0068] Preferably, the vitamin included in the food supplement is chosen from a group consisting of Vitamin B1, Vitamin B2, Vitamin B6, Vitamin E, Vitamin C, Vitamin B9, Vitamin B12 and Vitamin B5. Each of these vitamins plays a crucial role in various physiological processes, including those related to the functioning of the nervous system.

[0069] Preferably, the amino acid derivative included in the food supplement can be, for example, carnitine. Carnitine is involved in the transport of fatty acids in mitochondria, where they are degraded to generate energy. This process is particularly important in neurons, which have high energy demands.

[0070] Preferably, the precursor of the synthesis of membrane phospholipids included in the food supplement can be, for example, citicoline. Citicoline is involved in the synthesis of phosphatidylcholine, a main component of cell membranes. The inclusion of citicoline in the food supplement can support the integrity and function of the neuronal cell membranes, thus contributing to the health of the nervous system.

[0071] In some cases, the food supplement can be formulated as a tablet, capsule, powder or liquid, and can be taken by mouth. The specific form of the food supplement can depend on various factors, such as the intended use of the supplement, the user’s preferences and the stability of the components in different forms.

[0072] In one embodiment, the food supplement can be used in combination with other treatments or interventions, such as drugs, physiotherapy or changes in lifestyle.

[0073] In some aspects, the food supplement can be used as part of a daily regime to support nervous system health and function. The use of the food supplement, in this manner, can provide a proactive approach for maintaining neurological health and preventing the onset of neurological disorders.

[0074] In some cases, the food supplement can be used in populations at risk of neurological disorders, such as older adults or individuals with a family history of neurological disorders. The use of the food supplement in these populations can provide a potential strategy for reducing the risk of developing neurological disorders.

[0075] In some embodiments, the food supplement can be used in populations with existing neurological disorders, such as individuals diagnosed with Alzheimer’s disease, Parkinson’s disease, multiple sclerosis, neuropathic pain, epilepsy or stroke. The use of the food supplement in these populations can provide a potential approach for managing the symptoms of these disorders and improving the quality of life of these individuals. In some aspects, the food supplement can be used in populations with non-neurological conditions that can influence nervous system health and function, such as diabetes, cardiovascular diseases or chronic kidney diseases. The use of the food supplement in these populations can provide a potential approach for addressing the neurological complications associated with these conditions.

[0076] In some cases, the food supplement can be used in healthy populations to support nervous system health and function. The use of the food supplement in these populations can provide a proactive approach for maintaining neurological health and preventing the onset of neurological disorders.

[0077] In some embodiments, the food supplement can be used in populations with specific nutritional needs, such as athletes, pregnant women or individuals who follow a vegetarian or vegan diet. The use of the food supplement in these populations can provide a potential approach for satisfying the specific nutritional needs of these individuals and support the health and function of their nervous system.

[0078] In some aspects, the food supplement can be used in populations with specific lifestyle factors which can influence nervous system health and function, such as individuals who smoke, consume alcohol or experience high levels of stress. The use of the food supplement in these populations can provide a potential approach for mitigating the negative impact of these lifestyle factors on the nervous system.

[0079] In some cases, the food supplement can be used in populations with specific environmental exposures that can influence nervous system health and function, such as individuals exposed to toxins, pollutants or radiation. The use of the food supplement in these populations can provide a potential approach for mitigating the negative impact of these environmental exposures on the nervous system.

[0080] In some embodiments, the food supplement can be used in populations with specific genetic factors that can influence nervous system health and function, such as individuals with genetic mutations or polymorphisms associated with neurological disorders. The use of the food supplement in these populations can provide a potential approach for mitigating the negative impact of these genetic factors on the nervous system.

[0081] In some aspects, the food supplement can be used in populations with specific physiological factors that can influence nervous system health and function, such as individuals with hormone imbalances, immune dysregulation or metabolic disorders. The use of the food supplement in these populations can provide a potential approach for mitigating the negative impact of these physiological factors on the nervous system. In some cases, the food supplement can be used in populations with specific psychological factors that can influence nervous system health and function, such as individuals with mental health disorders, cognitive impairments or sleep disorders. The use of the food supplement in these populations can provide a potential approach for mitigating the negative impact of these psychological factors on the nervous system.

[0082] In some cases, the food supplement can be used in populations with specific geographic factors that can influence nervous system health and function, such as individuals who live in regions with high elevations, extreme temperatures or limited sunlight. The use of the food supplement in these populations can provide a potential approach for mitigating the negative impact of these geographic factors on the nervous system.

[0083] In some embodiments, the food supplement can be used in populations with specific occupational factors that can influence the nervous system health and function, such as individuals employed in jobs that entail heavy physical labour, high levels of stress or exposure to harmful substances. The use of the food supplement in these populations can provide a potential approach for mitigating the negative impact of these occupational factors on the nervous system.

[0084] EXAMPLE

[0085] The following substances were tested:

[0086] - Chaga 250 pg / mL;

[0087] - Chaga 1000 pg / mL;

[0088] - Carnitine 500 pg / mL;

[0089] - Carnitine 250 pg / mL;

[0090] - Citicoline

[0091] - Mix1 : Chaga 250 pg / mL+ Carnitine 500 pg / mL+Citicoline; and

[0092] - Mix2: Chaga 1000 pg / mL+ Carnitine 250 pg / mL+Citicoline

[0093]

[0094] The study was conducted using well-designed in vitro models called Transwell®, which provide for the use of human intestinal epithelial cells, the Caco-2 cell line, widely used and validated as a model for absorption studies on compounds administered orally.

[0095] The same 3D in vitro model (approved by the FDA and EMA) was used to test absorption and the mechanisms of transport across the intestinal barrier, by analysing:

[0096] - Cell viability by means of the MTT assay

[0097] - TEER measurement

[0098] - Tight junctions activity

[0099] The cells seeded in the Transwell® insert were maintained in a complete medium, changed every other day for 21 days prior to the stimulations, in order to ensure the maturation and formation of intestinal microvilli. The maturation was complete upon the reaching of a transepithelial resistance (TEER) value >400n cm2.

[0100] Analysis of safety at the level of the central nervous system

[0101] The product metabolised by the intestinal cells was placed directly in contact with astrocytes (GGF-STTG1). This technique was developed by simulating a condition of cognitive decline induced by H2O2 200 pM and analysing:

[0102] Survival and mitochondrial activity (viability and ROS)

[0103] The main pathways involved in cognitive decline processes, APP, pTAU and Sirt-1.

[0104] Results

[0105] At the intestinal level, all the substances are capable of improving intestinal viability compared to the control (p<0.05); specifically, the Mixes prove to be better than the single components (p<0.05) but Mix 2 appears to perform better than Mix 1 (p<0.05). (Figure 1).

[0106] The TEER analyses confirm the active role of the samples examined and their absorption. Furthermore, the analysis of tight junctions such as Zo-1 (which mediates adhesion); Claudin (which maintains the structure) and Occludin (which contributes to stabilisation) confirm correct intestinal function. None of the substances examined causes irritability at the intestinal level. (Figure 2 and 3).

[0107] Brain tissue treated with 200pM of H2O2 significantly reduced biological activity (p < 0.05 vs the control), indicating the start of the neuronal aging / degeneration cascade.

[0108] This negative condition was significantly countered by the presence of Mix 2, which showed a synergistic effect among the components, surpassing the effects of the single agents and of Mix 1 (p<0.05). The same behaviour was observed on oxidative stress, which was reduced following treatment with Mix 2 as opposed to the single agents and Mix 1 at the level of the CNS (p<0.05). (Figure 4).

[0109] Sirt-1 controls energy metabolism via the gluconeogenic / glycolytic pathways through PGC-1a, leading to an increase in mitochondrial function. As may be observed, all the agents examined improved the levels of Sirt-1 also after the damage induced by H2O2200 pM (p<0.05). Furthermore, the expression of APP (beta-amyloid precursor) and pTAU (cognitive decline) was analysed; as regards pTAU, the previously observed data concerning the beneficial effect on brain trophism was confirmed; the tested Mixes, specifically Mix 2, reduce the levels of pTAU with respect to the induced damage (p<0.05). Furthermore, the data emerging from the analysis of APP support the hypothesis of the prevention of cognitive decline; in fact, Mix 2 was capable of inducing a significant improvement with respect to the damaged induced by H2O2200 pM, compared to the single agents and Mix 1 (p<0.05). (Figure 5). To imitate the damage to peripheral nerve tissue in vitro, 3D EngNT was pretreated starting from the 14thday of maturation with 200 ng / mL of glial growth factor 2 (GGF) to reproduce a robust demyelination prior to stimulation with the same agents as used previously. In particular, the nerve tissue treated with only 200 ng / mL of GGF significantly reduced the biological activity of the nerve (p < 0.05 vs the control). In contrast, this negative condition was significantly countered by the presence of Mix 2, which showed a synergistic effect among the components, surpassing the effects of the single agents and of Mix 1 (p<0.05), and suggesting the possibility of repairing nerve damage. Furthermore, Mix 2 exerted a greater beneficial effect compared to the single agents and Mix 1 (p<0.05) in terms of oxidative stress, reducing ROS production at the level of the PNS. (Figure 6).

[0110] The role of Schwann cells was studied by analysing the modulation of neuropathic pain in vitro. After the nerve injury induced by GGF, the myelinated cells were subjected to degradation, as demonstrated by the analysis of p75 and, consequently, the inhibition of the activity of MPZ. In contrast, Mix 2 reduced the damage, thus confirming its positive role in countering the demyelination process, and exerted a greater beneficial activity compared to the single agents and Mix 1 (p<0.05). (Figure 7).

[0111] The role of Schwann cells was studied by analysing the modulation of neuropathic pain in vitro. The condition of PNI influenced the expression of neuregulin 1 (NRG1) and the activity of the epidermal receptor beta (ERb) (p < 0.05). In contrast, Mix 2 reduced the damage, thus confirming its positive role in countering the demyelination process and exerting a greater beneficial activity compared to the single agents and Mix 1 (p<0.05). (Figure 8).

[0112] Based on the previously obtained data, it was decided to evaluate the systemic effects of the two study Mixes. Specifically, the effects of the final formula on lipid metabolism were evaluated. Since cholesterol and triglycerides can increase during the aging process, additional experiments were conducted to analyse their levels:

[0113] Mix 2 showed a bioactive effect in modulating the levels of triglycerides and total cholesterol compared to the control (p<0.05).

[0114] Mix 2 was demonstrated to amplify the effects of the single agents (p<0.05), supporting the synergistic effect among the components (p<0.05).

[0115] As regards both triglycerides and cholesterol, Mix 2 showed a greater beneficial effect compared to Mix 1 (p<0.05). (Figure 9).

Claims

CLAIMS1. A composition comprising an extract of a fungus belonging to the family Hymenochaetaceae, at least one vitamin and / or carnitine for use in the treatment or prevention of a pathological condition of the central and / or peripheral nervous system, characterized in that the fungus belonging to the family Hymenochaetaceae is Inonotus obliquus (Chaga).

2. The composition for use according to claim 1 , wherein the extract of the fungus belonging to the family Hymenochaetaceae is obtained from the fruiting body of the fungus or from at least a portion thereof.

3. The composition for use according to claim 1 or 2, wherein the extract of the fungus belonging to the family Hymenochaetaceae is obtained by extraction in an aqueous solvent.

4. The composition for use according to any one of claims 1-3, wherein the at least one vitamin is chosen from: Vitamin B1 , Vitamin B2, Vitamin B6, Vitamin E, Vitamin C, Vitamin B9, Vitamin B12 and Vitamin B5.

5. The composition for use according to claim 4, comprising at least three vitamins.

6. The composition according to claim 5, wherein the composition comprises at least four vitamins chosen from: Vitamin B1, Vitamin B2, Vitamin B6, Vitamin E, Vitamin C, Vitamin B9, Vitamin B12, Vitamin B5.

7. The composition for use according to any one of claims 1-6,further comprising citicoline.

8. The composition for use according to any one of claims 1-7, wherein the pathological condition is selected from the group consisting ofAlzheimer's disease, Parkinson's disease, multiple sclerosis, neuropathic pain, epilepsy and stroke.

9. A food supplement comprising an extract of a fungus belonging to the family Hymenochaetaceae, at least one vitamin, and carnitine.

10. The food supplement according to claim 9, further comprising citicoline.

11. The food supplement according to claim 9 or 10, wherein the fungus belonging to the family Hymenochaetaceae is Inonotus obliquus (Chaga).

12. The food supplement according to any one of claims 9-11 , wherein the at least one vitamin is chosen from: Vitamin B1 , Vitamin B2, Vitamin B6, Vitamin E, Vitamin C, Vitamin B9, Vitamin B12, Vitamin B5.

13. The food supplement according to any one of claims 9-12, wherein the extract of the fungus belonging to the family Hymenochaetaceae is obtained from the fruiting body of the fungus or from at least a portion thereof.

14. The food supplement according to any one of claims 9-13, wherein the extract of the fungus belonging to the family Hymenochaetaceae is obtained by extraction in an aqueous solvent.

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